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"Impact of cap structures on the performance of in vitro-transcribed mRNAs"

Exploring strategies for optimal capping to maximize mRNA potency

Capping is a crucial modification of in vitro–transcribed mRNAs, enhancing their stability, facilitating translation initiation, and enabling self/non-self discrimination in therapeutic applications. At its most basic level, capping involves the addition of a 7-methylguanosine (m7G) to the 5′ end of the mRNA generating a Cap-0 structure, which is often recognized as non-self in higher eukaryotes. The first transcribed base can also be methylated at the 2′ ribose position to generate the more common Cap-1 structure found on endogenous mRNAs in higher eukaryotes. 

This tech note details a comprehensive evaluation of mRNA capping strategies, focusing on CleanCap® technology and its impact on mRNA quality and protein expression both in vitro and in vivo. 

We report the following in this tech note: 

• Evaluate mRNA quality using enzymatic and CleanCap capping methods 
• Present protein expression data for three CleanCap cap analogs 
• Compare in vivo protein expression of CleanCap M6 and enzymatic capping 
• Demonstrate the possibility of dose reduction with CleanCap M6 mRNA 

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